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. Author manuscript; available in PMC: 2011 Mar 12.
Published in final edited form as: Biochem Biophys Res Commun. 2010 Feb 10;393(3):504–508. doi: 10.1016/j.bbrc.2010.02.035

Fig. 2. In vitro P16 Phosphorylation.

Fig. 2

A, P16 phosphorylation by recombinant IKKβ. B, P16 phosphorylation by cell lysates and IKKβ-depleted cell lysates. WI38--IKKβ and U2OS--IKKβ represent IKKβ-depleted WI38 and U2OS lysates, respectively. Each reaction mixture contained 0.1 μg of recombinant IKKβ (A) or 10 μg of cell lysates (B), 2 μg of indicated proteins, and 5 μCi [γ-32P] ATP in a volume of 15 μl. In A, IκBα1-214 and Yar 1 were used as positive and negative controls, respectively. P16 SSSS, P16 WT; P16 AAAA, S7A/S8A/S140A/S152A; P16 SAAA, S8A/S140A/S152A; P16 ASAA, S7A/S140A/S152A; P16 AASA, S7A/S8A/S152A; P16 AAAS, S7A/S8A/S140A.